Ergot alkaloids are a complex family of indole derivatives with diverse structures and biological activities (Flieger 1997, Folia Microbiol (Praha) 42:3-30; Schardl 2006, Chem Biol 63:45-86) and are produced by fungi of the families Clavicipitaceae (e.g. Claviceps and Neotyphodium or Epichloee) and Trichocomaceae (including Aspergillus and Penicillium). Important natural producers are fungi of the genera Claviceps, Penicillium, and Aspergillus (Flieger 1997, loc cit.; Schradl loc cit.), but they are also found in the genus Sphacelia, Balansia or Periglandula (Pa{hacek over (z)}outová, S. et al, 2008, Fungal Diversity, 31: 95-110 and Steiner, U. 2011, Mycologia, 103(5):1133-1145). Both the natural ergot alkaloids and their semisynthetic derivatives are in widespread use in modern medicine and exhibit a broad spectrum of pharmacological activities, including uterotonic activity, modulation of blood pressure, control of the secretion of pituitary hormones, migraine prevention, and dopaminergic and neuroleptic activities (de Groot 1998, Drugs 56:523-535; Haarmann 2009, Mol Plant Pathol 10:563-577; Schardl 2006, loc cit.) Ergot alkaloids can be divided into two classes according to their structural features, i.e., amide derivatives of D-lysergic acid and the clavine-type alkaloids (Flieger 1997, loc cit.) Gröger 1998, Alkaloids Chem Biol 50:171-218). The members of the first group are usually composed of lysergic acid and a peptide moiety. They are also referred to as ergopeptines. Important members of the ergopeptines are for example ergotamine and ergovaline. The clavine-type alkaloids like cyloclavine, agroclavine, fumigaclavine and similar substances, like elymoclavine, pyroclavine, costaclavine or epcostaclavine as well as their precursors chanoclavine-I and chanoclavine aldehyde merely consist of the same or a similar tricyclic or tetracyclic ring system, like the ergopeptines, but lack a peptide moiety. However, clavine-type alkaloids may also comprise additional substitutents and are thus not limited to the clavine-type alkaloids mentioned above. Examples of such derivatives are the group of fumigaclavines, like fumiclavine A, B or C, isofumigaclavine A or B, or 9-deacetylfumigaclaine C (Wallwey, C. and Li, S. M. 2011, Nat. Prod. Rep., 28:496-510) share the basic structure with festuclavine, but comprise additional substituents. Fumigaclavines are for example produced by Penicillium and Aspergillus, e.g., A. fumigatus (Flieger 1997, loc cit.). The fungal family of the Clavicipitaceae e.g. C. purpurea (Flieger 1997, loccit), however have the capacity to produce ergopeptines, which can be considered to be derivatives of lysergic acid, of one of which precursors is the clavine-type alkaloid agroclavine. Comparison of the precursors of ergopeptidens and clavine-type alkaloids indicated that the early stages of their biosynthetic pathway are very similar and likely shared for example by A. fumigatus and C. purpurea, whereas later steps in the pathway differ in the two fungal species (Li 2006, Chembiochem 7:158-164; Panaccione 2005, FEMS Microbiol Lett 251:9-17; Schardl 2006, loc cit.).
The fermentative production of clavine-type alkaloids and even ergot alkaloids in general has to cope with the problem that natural producers of ergot alkaloids tend to be hard to culture in large scale. One reasons for this are technical problems of fermenting filamentous fungi in submerse culture. Some natural ergot alkaloid producers are even endophytes of plant genera like Ipomoea, Turbina, Argyreia and Strictocardia (e.g. Periglandula species) or of certain grass species, like Lolium, Sorghum or Festuca (e.g. Neotyphodium and Epichloe species). Further, many natural ergot alkaloid producers produce these alkaloids only under specific conditions or during certain developmental stages, even then they usually produce at a low production rate. Many natural ergot alkaloid producers produce a collection of different ergot alkaloids which causes problems to isolate a given alkaloid in a cost effective manner and lowers the production of a particular alkaloid of interest even further. The total chemical synthesis of ergot alkaloids or even of the less complex clavine-type alkaloids does, so far, still represent a considerable challenge and results usually in the synthesis of racemic mixtures.
In order to overcome these problems, the present invention provides recombinant microorganisms and recombinant natural ergot alkaloid producer organisms, as well as polynucleotides, polypeptides, vectors and methods using these recombinant microorganisms and recombinant natural ergot alkaloid producer organisms, as well as polynucleotides, polypeptides and vectors for the production of clavine-type alkaloids.